Construction of membrane-bound macrophage colony-stimulating factor and recombinant retroviral expression vector of spliceosome
MA Cui-hua
LIAO Jin-feng
WANG Da-gang
LIU Shu-yan
REN Qian
ZHENG Guo-guang
Abstract：Objective To construct membrane-bound macrophage colony-stimulating factor ( mM-CSF) and recombi-nant retroviral expression vector of brachytmema mutation of 30 amino acide located in the intracellular region of mM-CSF ( mM-CSF-Δ) .Methods The retroviral vectors MSCV-PGK-GFP-mM-CSF and MSCV-PGK-GFP-mM-CSF-Δwere con-structed and identified by DNA recombinant techniques.Recombinant and empty vectors were used to transfect the packa-ging Phoenix cells.HEK293 cells were infected by the viral supernatants.After being sorted by flow cytometry, three posi-tive cell lines were obtained.Results HEK293 cells were successfully infected by retroviruses in packaging Phoenix cells and control retrovirus.Stable expressing cell lines, HEK293-M and HEK293-M-Δas well as control cell line HEK293-V, were established.The expression of mM-SCF and mM-CSF-Δwas detected by RT-PCR and Western blotting in HEK293 cells and its membrane protein expression was also detected by flow cytometry.Conclusion The mM-CSF and recombinant retroviral expression vector of spliceosome were successfully constructed.
Keywords：membrane-bound macrophage colony-stimulating factorspliceosomeretroviral expression vectorHEK293 cell line
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 5-7 )